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ToxSci Advance Access originally published online on June 12, 2003
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Toxicological Sciences 75, 148-153 (2003)
Copyright © 2003 by the Society of Toxicology

REPRODUCTIVE AND DEVELOPMENTAL TOXICOLOGY

The Laminin Binding Protein p40 Is Involved in Inducing Limb Abnormality of Mouse Fetuses as the Effects of Methoxyacetic Acid Treatment

Aceng Ruyani*,1, Sri Sudarwati*, Lien A. Sutasurya*, Sony H. Sumarsono* and Torsten Gloe{dagger}

* Departemen Biologi, Institut Teknologi Bandung Jl, Ganesha No. 10, Labtek XI, Bandung 40132, Indonesia, and {dagger} Institute of Physiology, Ludwig Maximilians University, Schillerstrasse 44, D 80336 Munich, Germany

This study is intended to characterize a protein that is linked with mouse limb teratogenicity as the effects of methoxyacetic acid (MAA) treatment. A single dose of MAA (10 mmol/kg body weight) was given by gavage on gestation day (GD) 11, whereas the control group were administered vehicle only. The pregnant mice were killed at 4 h after MAA treatment, and forelimb buds were isolated from both the control and treated group embryos. Proteins from forelimb buds GD 11 + 4 h, which were precipitated out using 40–60% ammonium sulfate, then were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2-D SDS–PAGE) technique. The 2-D gels reveal one protein with 41.6 kDa and pI 6.4, which expression was downregulated after MAA treatment. Tentative protein identification via peptide mass database search and definitive protein identification via a primary sequence database search indicate that the protein matches exactly to 34/67 kDa laminin binding protein (LBP; P14206, SwissProt), which is encoded by p40 gene (MGI:105381). The identity was further verified by Western blotting with an antibody against the 67 kDa LBP. The results suggest that MAA treatment to pregnant mice downregulates the LBP-p40 in the forelimb buds.

Key Words: limb teratogenicity; methoxyacetic acid; 2-D SDS–PAGE; LBP-p40.


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